SN/T 5334.4-2020Digital PCR Detection Method for Genetically Modified Plant Products - Part 4: Genetically Modified Rape (English PDF)

Foreword

SN/T 5334-2020 "Digital PCR Detection Method for Genetically Modified Plant Products" is divided into 8 parts.

--Part 1: General requirements and definitions;

--Part 2: Genetically modified soybeans;

--Part 3: Genetically modified corn;

--Part 4: Genetically modified rape;

--Part 5: Genetically modified cotton;

--Part 6: Genetically Modified Potatoes;

--Part 7: Genetically modified alfalfa;

--Part 8: Genetically modified sugar beet.

This document is part 4 of SN/T 5334-2020.

This document was drafted in accordance with the rules given in GB/T 1.1-2020.

Certain contents of this document may involve patents. The issuing agency of this document is not responsible for identifying these patents.

This document was proposed and managed by the General Administration of Customs of the People's Republic of China.

Drafting organizations of this document. Chinese Academy of Inspection and Quarantine Sciences, Shenzhen Customs Food Inspection and Quarantine Technology Center of the People's Republic of China, Shanghai Customs of the People's Republic of China, Gongbei Customs Inspection and Quarantine Technology Center of the People's Republic of China.

1 Scope

This document specifies the digital PCR quantitative detection method for genetically modified strains in rapeseed.

This document is applicable to the quantitative detection of 11 genetically modified lines in rapeseeds and leaves. See Appendix A for the detection limits of this document.

2 Normative references

The contents of the following documents constitute the indispensable clauses of this document through normative references in the text. Among them, dated quotations Only the version corresponding to that date is applicable to this document; for undated references, the latest version (including all amendments) is applicable Used in this document.

SN/T 5334.1 Digital PCR Method for Quantitative Detection of Genetically Modified Products Part 1: General Requirements and Definitions

3 Terms and definitions

The terms and definitions defined in SN/T 5334.1 apply to this document.

4 Principle

See SN/T 5334.1.

5 Equipment and reagents

5.1 Endogenous gene and exogenous gene. the internal standard gene is the rape phosphoenolpyruvate carboxylase gene (PEP); the exogenous sequence is the product Lines 73496, T45, MON88302 and OXY235 foreign gene insertion sites 5'end cross-border sequence, and lines Topas19/2, MS1, the foreign sequences of MS8, Rf1, Rf2, Rf3 and RT73 are inserted into the cross-border sequence at the 3'end of the site. See Appendix A for primer and probe sequences.

5.2 For other equipment and reagent requirements, please refer to SN/T 5334.1.

6 Method operation steps

7 Judgment and expression of results

See SN/T 5334.1 for calculation of results.

The results are expressed as follows.

- In the test results, both the internal standard gene and the foreign gene meet the negative and blank quality control requirements, indicating that no rape has been detected.

Source gene PEP, expressed as "No oilseed rape component detected";

- In the test results, the internal standard gene result is positive, but the foreign gene meets the quality control requirements of negative and blank, said Genetically modified strains";

- In the test results, both the internal standard gene and the exogenous genome meet the quality control requirements of the digital PCR test of the sample, indicating that

8 Anti-pollution measures

The anti-pollution measures are specified in SN/T 5334.1.

9 Sample storage

For sample storage, see SN/T 5334.1.

Appendix A

(Informative appendix) Primer probe sequence, reaction program and detection limit